摘要
目的:研究不同中医辨证分型晚期肺癌患者血清microRNA(miRNA)表达谱差异。方法:将经病理诊断明确为非小细胞肺癌患者共9例,并按肺癌中医证型诊断标准分为气阴两虚型3例、脾虚痰湿型3例、气滞血瘀型3例,以同期健康体检者3例作为对照组,应用高通量测序法检测各组血清中miRNA的表达谱,筛选出差异表达的miRNA,实时荧光定量PCR验证结果,并进行靶基因预测及生物信息学分析。结果:在健康对照组及气阴两虚型、脾虚痰湿型、气滞血瘀型患者中分别鉴定得到791、707、773、846个已知miRNA成熟体。通过miRNA差异分析,发现hsa-miR-103a-3p、hsa-miR-103b、hsa-miR-139-5p、hsa-miR-532-5p等4个miRNA仅在气阴两虚型患者血清中呈显著表达;hsa-miR-142-3p、hsa-miR-15b-5p、hsa-miR-16-5p、hsa-miR-18a-3p等15个miRNA仅在气滞血虚型患者血清中显著表达;hsa-let-7d-5p、hsa-miR-106b-5p、hsa-miR-107、hsa-miR-17-5p等18个miRNA仅在脾虚痰湿型患者血清中显著表达。实时荧光定量PCR对其中部分特异表达的miRNA验证结果与高通量检测结果一致。生物信息学分析发现,差异表达的miRNA调控的靶基因主要参与细胞周期、细胞凋亡、细胞信号传导、生物调控等生物学过程。结论:不同中医辨证分型肺癌患者血清miRNA表达谱存在明显差异,提示miRNA可能通过靶基因在不同中医辨证分型肺癌患者发病过程中发挥不同的调节作用。
Objective:To study the differences in microRNA(miRNA)expression profile of serum in patients with nonsmall cell lung cancer of different Chinese medicine syndrome types.Methods:A total of nine patients diagnosed with nonsmall cell lung cancer by pathology were divided into three groups according to the diagnostic criteria of Chinese medicine syndrome type of lung cancer,including patients of qi and yin deficiency type,patients of spleen deficiency and phlegmdampness type,and patients of qi stagnation and blood stasis type.Three healthy physical examinees were selected as the control group.The miRNA expression profile of serum in each group were detected by high-throughput sequencing,and the differentially expressed miRNAs were screened out.The results were verified by real-time fluorescence quantitative PCR,and the target genes were predicted and bioinformatics analyzed.Results:A total of 791,707,773 and 846 mature miRNA were respectively identified in the healthy control group,patients of qi and yin deficiency type,patients of spleen deficiency and phlegm-dampness type,and patients of qi stagnation and blood stasis type.Through difference analysis among miRNAs,it was found that four miRNAs,including hsa-miR-103 a-3 p,hsa-miR-103 b,hsa-miR-139-5 p and hsa-miR-532-5 p,were only significantly expressed in the serum of patients of qi and yin deficiency type;fifteen miRNAs,such as hsa-miR-142-3 p,hsa-miR-15 b-5 p,hsa-miR-16-5 p,hsa-miR-18 a-3 p,etc.,were only significantly expressed in the serum of patients of qi stagnation and blood stasis type;eighteen miRNAs,such as hsa-let-7 d-5 p,hsa-miR-106 b-5 p,hsa-miR-107 and hsa-miR-17-5 p,etc.,were only significantly expressed in the serum of patients of spleen deficiency and phlegm-dampness type.The results of real-time fluorescence quantitative PCR were consistent with those of highthroughput sequencing.Bioinformatics analysis revealed that target genes regulated by differentially expressed miRNA were mainly involved in biological processes such as cell cycle,apoptosis,cell signaling,and biological regulation.Conclusion:There are significant differences in the microRNA expression profile of serum in patients of different Chinese medicine syndrome types of lung cancer,indicating that miRNA may play different regulatory roles in the pathogenesis of patients through target genes.
作者
陈观平
汪一帆
应栩华
CHEN Guanping;WANG Yifan;YING Yuhua
出处
《新中医》
CAS
2020年第16期46-50,共5页
New Chinese Medicine
基金
浙江省科技计划资助项目(2016F10025)
浙江省中医药科技计划资助项目(2017ZB017)。
关键词
非小细胞肺癌
辨证分型
MIRNA
高通量筛选
Non-small cell lung cancer
Syndrome differentiation type
MiRNA
High-throughput screening
